JP2020514708A - Methods for tracking sample identity during processing in analytical systems - Google Patents

Abstract

A method for tracking sample identity during a process in an analytical system (112) is disclosed. The analytical system (112) includes a liquid chromatograph (102) and a mass spectrometer (104). The method provides a sample (108), adds a composition (130) of different chemicals (132) to the sample (108) at a concentration above the detection level of the mass spectrometer (104), and 108) with the composition (130) are processed by the analytical system (112), and one or more components of the sample (108) are detected and measured by the mass spectrometer (104) to detect different chemistries. Substance (132) composition (130) or portion of composition (130) detected by mass spectrometer (104) and measured, and substance corresponding to composition (130) of chemical substance (132) Identifying the sample (108) based on the detection of the substance detection signal pattern of the mass spectrometer (104) including the detection signal.

Description

  A method for tracking sample identity during a process in an analytical system is disclosed.

  There is an increasing interest in performing mass spectrometry, and more specifically liquid chromatography coupled with tandem mass spectrometry (LC-MS / MS) in different types of laboratories, such as clinical laboratories. In particular, the number of published methods is increasing for small molecules in therapeutic drug surveillance or drug abuse testing.

  Several ready-to-use kits are becoming commercially available for pre-validated clinical MS applications. However, the use of mass spectrometry, even in combination with such kits, may not be approved by regulatory authorities for clinical diagnosis. This is largely because there is no standardized method except for very few analytes, and, for example, there are still many manual steps involved and are both usable and combinable, and Due to the variety of hardware components that play a role in delivering reliable and reproducible results of clinical relevance, there are still many user-dependent factors. In particular, sample preparation is typically a manual and tedious procedure. Subsequent protein precipitation with centrifugation is the most common method to remove unwanted and potentially interfering sample matrix. Use of the kit may facilitate, at least in part, automatable sample preparation. However, kits are only available for a limited number of analytes of interest, and the entire process from sample preparation to separation and detection remains complex, making it extremely sophisticated to handle equipment. Requires the presence of highly trained laboratory personnel.

  Also typically, according to the batch approach, batches of pre-prepared sample under the same preparation conditions undergo continuous separation runs under the same separation conditions. However, this approach is not capable of high throughput and is inflexible, for example with higher priority, and rescheduling (pre-defined) to account for incoming urgent samples that need to be processed first. Processing sequence changes).

  Performing liquid chromatography coupled with tandem mass spectrometry allows several samples to be processed in parallel. Ensuring traceability of sample identity during complex and highly collimated random access sample preparation processes is an important issue. This is especially important in a multiplexed HPLC-MS system where many samples are processed in parallel at the same time and in part with the same liquid flow path. In such cases, sample identity is only obtained by the correct timing and placement of the diversion valves that control the flow path, which considerably complicates sample handling.

  Aspects of the disclosed methods provide for better and more reliable traceability of sample identity in analytical systems using liquid chromatography coupled with mass spectrometry, and thus specific analytical methods such as clinical. The purpose is to make it suitable for diagnosis. In particular, random access sample preparation and LC separation may provide high throughput, eg, up to 100 samples / hour or more, while each sample is traceable throughout the process. Furthermore, the method is fully automatable, it is possible to increase the walk-away time and to reduce the required technical level.

Disclosed herein is a method for tracking sample identity during a process in an analytical system, wherein the analyzer comprises a liquid chromatograph and a mass spectrometer.
Aspects of the disclosed methods for tracking sample identity during processes in analytical systems have the features of independent claims. Further aspects of the invention, which can be realized alone or in any arbitrary combination, are disclosed in the dependent claims.

  The terms “have,” “comprise,” or “include” or any grammatical variations thereof, as used below, are used in a non-exclusive manner. Thus, these terms refer to the features introduced by these terms in addition to the circumstances in which there are no additional features present in the entity described in this context, and where one or more additional features are present. You may point both. For example, the expressions "A has B," "A includes B," and "A includes B" mean that in addition to B, there are no other elements in A (ie, A is exclusively). And consisting exclusively of B), as well as situations in which in addition to B there are one or more further elements in entity A, such as element C, elements C and D or even further elements. ..

  Further, the terms "at least one", "one or more", or similar expressions indicating that a feature or element may be present more than once or typically more than once It will be used only once in introducing the feature or element. After that, in most cases, when referring to each feature or element, the expression "at least one" or irrespective of the fact that each feature or element may occur once or more than once "One or more" will not be repeated.

  Further, as used below, the terms "particularly", "more specifically", "specifically", "more specifically" or similar terms. Are used in combination with additional / different features without limiting alternative possibilities. Therefore, the features introduced by these terms are additional / alternate features and are not intended to limit the scope of the claims in any way. The present invention can be implemented by using alternative features, as will be appreciated by those skilled in the art. Similarly, features introduced by "in an aspect of the invention" or similar phrases do not imply any limitation as to alternative aspects of the invention, as to the scope of the invention, and other aspects of the invention. Additional / different features are intended, without any limitation as to the possibility of combining features introduced in such a manner with additional / different or additional / non-different features.

According to the disclosed method, there is disclosed a method for tracking sample identity during a process in an analytical system, wherein the analytical system comprises a liquid chromatograph and a mass spectrometer. The method is:
-Providing a sample,
-Adding different chemical compositions to the sample at concentrations above the detection level of the mass spectrometer,
-Processing the sample with the composition by the analytical system,
-Detecting and / or measuring one or more components of the sample by a mass spectrometer,
-Compositions or parts of compositions of different chemicals detected and measured by mass spectrometry,
-Identifying the sample on the basis of the detection of the substance detection signal pattern of the mass spectrometer, which contains the substance detection signal corresponding to the composition of the chemical substance.

  The term “analytical system” as used herein includes any system set up to obtain a measurement, including at least a liquid chromatograph and a mass spectrometer. The analytical system is operable to determine the parameter values of the sample or its constituents through various chemical, biological, physical, optical or other technical methods. The analytical system may be operable to measure said parameter of the sample or at least one analyte and return the measured value obtained. The list of possible analytical results returned by the analyzer includes, without limitation, the concentration of the analyte in the sample, a digital (with or without) presence of the analyte in the sample (corresponding to concentrations above the detection level). ), Optical parameters, DNA or RNA sequences, data obtained from mass spectrometry of proteins or metabolites, and various types of physical or chemical parameters. The analytical system may include units that aid in pipetting, dosing, and mixing of samples and / or reagents. The analytical system may include a reagent holding unit for holding reagents to carry out the assay. The reagents may be arranged, for example, in the form of containers or cassettes containing individual reagents or groups of reagents, placed in suitable containers or locations in a storage compartment or conveyor. The analytical system may include a consumable feeding unit. Analytical systems may include process and detection systems whose workflow is optimized for a particular type of analyte. Examples of such analysis systems are clinical chemistry analyzers, coagulation chemistry analyzers used to detect the results of chemical or biological reactions or to monitor the progress of chemical or biological reactions. , An immunochemical analyzer, a urine analyzer, and a nucleic acid analyzer.

The analytical system may be designed as a clinical diagnostic system or may be part of a clinical diagnostic system.
"Clinical diagnostic system", as used herein, refers to a laboratory automation device that is specialized for the analysis of samples for in vitro diagnosis. The clinical diagnostic system may have different structures according to need and / or according to desired laboratory workflow. Further structures can be obtained by coupling multiple devices and / or modules together. A "module" is a working cell that is typically smaller in size than the entire clinical diagnostic system with specialized functions. This function may be an analytical function, but may also be a pre-analysis or post-analysis function, or it may be an adjunct to either a pre-analysis function, an analysis function, or a post-analysis function. It may be a function. In particular, the module is one or more for performing specialized tasks of a sample processing workflow, for example by performing one or more pre-analytical and / or analytical and / or post-analytical steps. It may be configured to work with more other modules. In particular, clinical diagnostic systems are designed to perform their respective workflows optimized for specific types of analysis, such as clinical chemistry, immunochemistry, coagulation, hematology, liquid chromatography separations, mass spectrometry, etc. It may also include one or more analytical devices. Thus, the clinical diagnostic system may include one analyzer or a combination of any of these analyzers with each workflow, where the pre-analytical and / or post-analytical modules are coupled to individual analyzers. It may be ringed or shared by multiple analytical devices. Alternatively, the pre-analysis function and / or the post-analysis function may be performed by a unit incorporated in the analyzer. The clinical diagnostic system is a functional unit, eg a liquid handling unit for pipetting and / or pumping and / or mixing liquids of samples and / or reagents and / or systems, and also sorting, storage and transport. It may also include functional units for identifying, separating, and detecting.

  The term "sample" is herein assumed to contain one or more analytes of interest, and whose qualitative and / or quantitative detection is associated with a clinical condition. Refers to a substance. Samples include any biological source such as blood, saliva, ocular lens fluid, cerebrospinal fluid, sweat, urine, breast milk, ascites, mucus, synovial fluid, peritoneal fluid, amniotic fluid, tissue, cells, etc. , May be derived from physiological fluids. The sample may be pre-treated before use, for example, plasma preparation from blood, dilution of mucus, lysis, etc .; treatment methods include filtration, centrifugation, distillation, concentration, no interference components. Activation and addition of reagents may be involved. In some cases, the sample as obtained from the source may be used directly, or after a pretreatment and / or sample preparation workflow to modify sample properties, such as after addition of an internal standard, another After dilution with a solution, or for example to allow the performance of one or more in vitro diagnostic tests or to concentrate (extract / separate / concentrate) the analyte of interest, and / or Alternatively, it may be used after mixing with reagents to remove matrix components that potentially interfere with the detection of the analyte (s) of interest. The term "sample" is used tend to refer to the sample before sample preparation, while the term "prepared sample" is used to refer to the sample after sample preparation. In non-specific examples, the term "sample" can also generally refer to either a sample before sample preparation, a sample after sample preparation, or both. Examples of analytes of interest are generally vitamin D, drugs of abuse, therapeutic agents, hormones, and metabolites. However, this list is not comprehensive.

  The term "liquid chromatograph", as used herein, refers to a device configured for use in liquid chromatography (LC). LC is a separation technique in which the mobile phase is a liquid. LC may be performed either in columns or in planes. Current liquid chromatography, which generally utilizes very small packed particles and relatively high pressure, is referred to as high performance liquid chromatography (HPLC). In HPLC, a sample is forced to pass through a column packed with a stationary phase composed of irregular or spherical particles, a porous integrated layer, or a porous membrane by a high-pressure liquid (mobile phase). Be done. HPLC has historically been divided into two different subclasses based on the polarity of the mobile and stationary phases. The method where the stationary phase is more polar than the mobile phase (eg toluene as mobile phase, silica as stationary phase) is referred to as normal phase liquid chromatography (NPLC) and the reverse (eg water-methanol mixture as mobile phase and stationary phase). C18 = octadecylsilyl) as the phase is called reverse phase liquid chromatography (RPLC).

  The term "mass spectrometer" as used herein refers to an instrument configured for use in mass spectrometry (MS). MS is an analytical technique that ionizes species and sorts ions based on mass-to-charge ratio. In simpler terms, mass spectra measure mass within a sample. Mass spectrometry is used in many different fields and is applied to complex mixtures with pure samples. Mass spectra are plots of ion signal as a function of mass to charge ratio. These spectra are used to determine the elemental or isotopic signatures of samples, the masses of particles and molecules, and to elucidate the chemical structure of molecules such as peptides and other compounds. In a typical MS method, a sample, which may be a solid, liquid, or gas, is ionized, for example by irradiating the sample with electrons. This can cause destruction of the sample molecule into some charged fragments. These ions are then separated according to their mass-to-charge ratio, typically by accelerating them and subjecting them to an electric or magnetic field: ions of the same mass-to-charge ratio produce the same amount of deflection. Will pass. Ions are detected by a mechanism capable of detecting charged particles, such as an electron multiplier. The results are displayed as a spectrum of the relative abundance of detected ions as a function of mass-to-charge ratio. Atoms or molecules in a sample can be identified by correlating known masses with identified masses or through a characteristic fragmentation pattern.

  The term "composition of different chemicals" as used herein refers to a mixture of different chemicals. The basic requirement is that the chemicals used have to follow the same workflow as the analytes, thus exhibiting nearly identical behavior during sample processing and chromatography compared to the analyte of interest. This can be accomplished by using a chemical isotope or substance that has a similar polarity to the analyte.

  The disclosed methods are used to provide potential for chemical coding. The possibility of chemical coding is a special feature made possible by the high multiplexing power of mass spectrometric detection, which allows the detection of further signals at any point during the measurement. The combination of several markers (presence / absence; high / low concentration etc.) will allow unambiguous labeling of all samples in the system.

Compositions of different chemicals may be added to the sample before, during, or after preparation of the sample.
Thus, the analytical system may include a sample preparation station for automated preparation of samples. A "sample preparation station" performs a series of sample processing steps aimed at removing or at least reducing interfering matrix components in a sample and / or enriching an analyte of interest in a sample. A pre-analytical module coupled to one or more analyzers or units in an analyzer designed to. Such processing steps may include any one or more of the following processing operations performed on a sample or samples in a serial, parallel, or alternating fashion: Liquid pipetting (aspiration and / or dispensing), liquid pumping, mixing with reagents, incubation at specific temperatures, heating or cooling, centrifugation, separation, filtration, sieving, drying, washing, resuspension, aliquots , Transfer, storage etc.

  A "reagent" is a sample, for example, to prepare a sample for analysis, allow a reaction to occur, or allow detection of physical parameters of the sample or the analyte contained in the sample. It is a substance used in the treatment of. In particular, the reagent is a reactant, eg, one that is capable of binding to, or chemically converting, one or more analytes present in the sample or unwanted matrix components of the sample, typically May be a compound or agent or a substance containing such reactants. Examples of reactants are enzymes, enzyme substrates, conjugated dyes, protein binding molecules, ligands, nucleic acid binding molecules, antibodies, chelating agents, promoters, inhibitors, epitopes, antigens and the like. However, the term reagent is used to include any liquid that can be added to a sample, including diluents containing water or other solvents or buffer solutions, or analytes on proteins, bound proteins or surfaces. Included are substances used for the disruption of specific or non-specific binding of. The sample may be provided in, for example, a sample container, such as a sample tube including primary tubes and secondary tubes, or a multi-well plate, or any other sample holding support. The reagents may be arranged, for example, in the form of containers or cassettes containing individual reagents or groups of reagents, and may be placed in suitable containers or locations within a storage compartment or conveyor. Liquids of other types of reagents or systems may be provided in bulk containers or through line supplies.

  For sample tracking, the combination of coding agents is added to the sample early in the sample preparation process, ideally immediately after dispensing an aliquot of the sample (patient sample aliquot) to the first sample processing vial.

  It is also possible to design the chemistry to prevent its retention on the stationary phase of a liquid chromatograph. Thus, the sample identity is clearly detectable and ensures that the chemical tracks the sample throughout the workflow so that the mass spectrometer does not detect the chemical during normal application.

For example, the polarity and molecular weight of the chemical is adapted to the stationary phase of the liquid chromatograph so as to prevent the chemical from being retained on the stationary phase of the liquid chromatograph.
The polarity of a chemical may be adapted by using a mixture of isobaric oligomers or isotopes of the chemical. Therefore, the chemicals used behave almost identically to the analyte of interest during sample processing and chromatography. Thereby, the identity of the sample can be unambiguously detected.

Molecular weights may be adapted by using homologues, derivatives or isotopic labels of chemicals. Therefore, the molecular weight can be adapted in a fairly simple manner.
The polarity of the chemical may be adapted by using a mixture of oligomers or derivatives of the chemical. Therefore, the polarity can be adapted in a fairly simple manner.

  Each oligomeric component can also have a labeling group for molecular weight coding, which labeling group can be measured intact or released during the mass spectrometry process and the mass spectrometer. Measured by In the first option, the labeling group remains intact in the backbone and the total mass is detected, which is possible if the backbone mass of each single component is isobaric. .. In the latter option, the labeling group is released or separated and its mass is detected, for example, in the ion source or collision cell of a mass spectrometer. In this option, the backbone masses need not be isobaric.

  The sample contains at least one analyte of interest, where the chemical is a chemical isotope, and / or has a polarity similar to the analyte of interest. Therefore, the chemical behaves almost identically to the analyte of interest during sample processing and chromatography. Therefore, the identity of the analyte is clearly traceable.

The chemical may include polar molecular groups and lipophilic molecular groups. Thus, the polarities may be individually adapted to the analyte of interest.
The chemical substance is (polar) _n- (lipophilic) _m , where (polar) represents a polar molecular group, (lipophilic) represents a lipophilic molecular group, and n and m are integers greater than 0. It can also be formed from a monomer block having a certain structure. Thus, the monomers may be designed to have the same molecular weight (isobaric building blocks), so that the overall molecular weight of the oligomeric subunits depends on the ratio n / m that produces the isobaric oligomeric backbone. Is independent of and constant.

For example, a chemical substance is a monomer building block that randomly polymerizes, n ^* (polar) and m ^* (lipophilic), in which (polar) indicates a polar molecular group and (lipophilic) indicates a lipophilic molecule. Group, wherein n and m are integers greater than 0. Therefore, the n and m sequence distributions are random depending on the desired readout of the mass spectrometer, the molecular weight only by using selected ion monitoring or by the molecular weight readout, and the sequence by the fragmentation pattern (daughter ion scan). It may or may not be random.

  The liquid chromatograph may include a liquid chromatographic separation station that includes a plurality of liquid chromatographic channels. The method further comprises providing a plurality of samples and adding a composition of different chemicals to each of the plurality of samples, wherein the compositions added to the plurality of samples are different from one another A plurality of samples based on the detection of the substance detection signal pattern of the mass spectrometer, including the substance detection signals that are processed by the analytical system and that correspond to different compositions of the chemicals.

  The term "liquid chromatographic separation station" is used herein to describe, for example, the matrix composition remaining after sample separation that may still interfere with the analyte of interest, such as matrix constituents, such as subsequent detection, e. An analytical device or a module in an analytical device designed to be subjected to a chromatographic separation of the prepared sample in order to separate it from the elements and / or to separate the analytes of interest from each other to allow individual detection or Refers to a unit. According to one embodiment, the liquid chromatographic separation station is a module in an intermediate analyzer or analyzer designed to prepare a sample for mass spectrometry and / or transfer the prepared sample to a mass spectrometer. Or a unit. In particular, the liquid chromatographic separation station is a multi-channel liquid chromatographic separation station including a plurality of liquid chromatography channels.

  A “liquid chromatography channel” comprises a stationary phase selected according to the sample (s) and the type of analyte, and, for example, according to polarity or log P value, size or affinity, as is generally known. At least one capillary tubing and / or pumping a mobile phase through a stationary phase to capture and / or separate and elute and / or transfer an analyte of interest under selected conditions. A fluid line including a liquid chromatography column. At least one liquid chromatography column in the at least one liquid chromatography channel may be replaceable. In particular, liquid chromatography may include more liquid chromatography columns than liquid chromatography channels, in which case multiple liquid chromatography columns may be exchangeably coupled to the same liquid chromatography channel. Capillary tubing may bypass the liquid chromatography column or may allow for dead volume adjustments to fine tune the elution time window.

  Liquid chromatographic separation stations also typically further include a sufficient number of pumps, such as a binary pump if conditions require the use of an elution gradient, and some switching valves.

  The number of compositions can also correspond to the number of liquid chromatography channels. Thus, for each channel, a chemical coding is provided that allows unambiguous tracking of the sample throughout the process.

The chemicals can also have the same molecular weight. Therefore, it is ensured that the behavior of the analyte of interest in the mass spectrometer is almost the same.
The chemical entity can also be at least one element selected from the group consisting of: peptides, derivatives of peptides, peptide nucleic acids, oligonucleotides, and oligomers. Therefore, it is also possible to use substances that can be handled well.

  Each chemical can also include isobaric oligomeric groups and at least one molecular weight coding group, where the isobaric oligomeric groups and molecular weight coding groups of the different chemicals are different from each other. Thus, it is possible to enable the production of diverse coding substances with different molecular weights. This is produced, for example, by using different amounts of stable isotope labels or different chemical functionalities within the molecular weight code. The function of the oligomer building blocks is to produce polar leaders with different chromatographic properties (logP) of the subunits but the same molecular weight. Thus, the combination of the molecular weight code and the isobaric oligomer backbone makes it possible to generate chemical coding entities.

  The invention, in one or more of the aspects contained herein, further comprises computer-executable instructions for performing the method according to the invention when the program is executed on a computer or computer network. Discloses and proposes a computer program including. In particular, the computer program may be stored on a computer-readable data carrier. Thus, in particular, one, more than one, or even all of the method steps as indicated above may be performed by using a computer or computer network, preferably by using computer scripts.

  The invention, in one or more of the aspects contained herein, further comprises program code means for carrying out the method according to the invention when the program is run on a computer or computer network. Discloses and proposes a computer program product having. In particular, the program code means may be stored on a computer readable data carrier.

  Furthermore, the invention relates to a data carrier, on which a data structure is stored, which has been loaded in a computer or a computer network, for example in a working memory or main memory of the computer or computer network, and which is then used herein. According to one or more of the aspects disclosed in, a data carrier is disclosed and proposed in which a method can be performed.

  The invention is further stored on a machine-readable carrier for performing the method according to one or more of the aspects disclosed herein when the program is executed on a computer or computer network. A computer program product having a program code means is proposed and disclosed. As used herein, a computer program product refers to a program as a tradable product. The product may generally be in any form, for example in paper form, or on a computer-readable data carrier. In particular, the computer program product may be distributed over a data network.

  Finally, the invention is modified, containing instructions readable by a computer system or computer network, for performing the method in accordance with one or more of the aspects disclosed herein. Propose and disclose data signals.

  Preferably, when referring to computer-implemented aspects of the invention, one or more of the method steps, or further method steps, of a method according to one or more of the aspects disclosed herein. Can all be performed by using a computer or computer network. Thus, generally, any of the method steps, including providing and / or manipulating data, can be performed by using a computer or computer network. In general, any of these method steps typically requires manual effort, except for certain aspects that perform sample provision and / or actual measurements. It can also be included.

In particular, the invention further comprises:
A computer or computer network comprising at least one processor, said processor being adapted to perform the method according to one of the aspects described herein.
A computer loadable data structure, the data structure being adapted to perform a method according to one of the aspects described herein while the data structure is being executed on a computer;
A computer script, wherein the computer program, while being executed on a computer, is adapted to carry out a method according to one of the aspects described herein.
A computer program comprising program means for performing the method according to one of the aspects described herein, while the computer program is running on a computer or computer network.
A computer program comprising program means according to the preceding aspect, wherein the program means is stored on a computer readable storage medium,
-A storage medium, the aspects described herein after the data structure is stored on the storage medium and the data structure is loaded into the main memory and / or the working memory of a computer or computer network. A storage medium, and a computer program product having program code means adapted to carry out a method according to one of the above, wherein the program code means is executed on a computer or computer network. , Discloses a computer program product, wherein program code means can be or is stored on a storage medium for performing the method according to one of the aspects described herein. To do.

The findings of the disclosed methods are summarized to disclose the following aspects:
Embodiment 1: A method for tracking sample identity during a process in an analytical system, the analytical system comprising a liquid chromatograph and a mass spectrometer, the method comprising:
-Providing a sample,
-Adding different chemical compositions to the sample at concentrations above the detection level of the mass spectrometer,
-Processing the sample with the composition by the analytical system,
-Detecting and / or measuring one or more components of the sample by a mass spectrometer,
-Compositions or parts of compositions of different chemicals detected and measured by mass spectrometry,
Said method comprising the step of identifying the sample based on the detection of the substance detection signal pattern of the mass spectrometer, which comprises the substance detection signal corresponding to the composition of the chemical substance.

Aspect 2: The method according to aspect 1, wherein the composition of the different chemicals is added to the sample before, during or after the preparation of the sample.
Aspect 3: The method according to Aspect 1 or 2, wherein the chemical is designed to prevent the chemical from being retained on the stationary phase of the liquid chromatograph.

  Aspect 4: The method according to aspect 3, wherein the polarity and molecular weight of the chemical is adapted to the stationary phase of the liquid chromatograph so as to prevent the chemical from being retained on the stationary phase of the liquid chromatograph. ..

Aspect 5: The method of aspect 4, wherein the polarity of the chemical is adapted by using a mixture of isobaric oligomers or isotopes of the chemical.
Aspect 6: The method according to aspect 3 or 4, wherein the molecular weight is adapted by using homologues, derivatives or isotopic labels of the chemical substances.

Aspect 7: The method of aspect 4, wherein the polarity of the chemical is adapted by using a mixture of oligomers or derivatives of the chemical.
Aspect 8: Each oligomeric component has a labeling group for molecular weight coding, the labeling group being measured intact or released during the mass spectrometry process and measured by a mass spectrometer. The method according to embodiment 7.

  Aspect 9: Any one of aspects 1-8, wherein the sample comprises at least one analyte of interest, the chemical is a chemical isotope, and / or has a polarity similar to the analyte of interest. The method described.

Aspect 10: The method of any one of aspects 1-9, wherein the chemical comprises polar and lipophilic molecular groups.
Aspect 11: The chemical substance is (polar) n- (lipophilic) m, where (polar) indicates a polar molecular group, (lipophilic) indicates a lipophilic molecular group, and n and m are 0 or less. 11. The method according to embodiment 10, wherein the method is formed from a monomer block having a structure of a large integer.

Aspect 12: A chemical substance is a monomer building block that polymerizes at random, n ^* (polar) and m ^* (lipophilic), in which (polar) represents a polar molecular group and (lipophilic) is lipophilic. 11. The method according to embodiment 10, wherein the molecular group is represented and n, m are integers greater than 0.

  Aspect 13: The method according to any one of aspects 1 to 12, wherein the liquid chromatograph comprises a liquid chromatography separation station comprising a plurality of liquid chromatography channels, the method further providing a plurality of samples, Compositions of different chemicals are added to each of the plurality of samples, wherein the compositions added to the plurality of samples are different from each other, the plurality of samples along with the composition of the chemicals are processed by the analytical system, and A method as defined above, which comprises the step of identifying each of the plurality of samples based on the detection of the substance detection signal pattern of the mass spectrometer, which comprises substance detection signals corresponding to different compositions.

Aspect 14: The method of aspect 13, wherein the number of compositions corresponds to the number of liquid chromatography channels.
Aspect 15: The method of any one of aspects 1-14, wherein the chemicals have the same molecular weight.

  Embodiment 16: The method according to any one of Embodiments 1 to 15, wherein the chemical substance is at least one element selected from the group consisting of: peptides, derivatives of peptides, peptide nucleic acids, oligonucleotides, and oligomers.

  Embodiment 17: Any of embodiments 1-16, wherein each chemical entity comprises an isobaric oligomeric group and at least one molecular weight coding group, wherein the isobaric oligomeric group and molecular weight coding group of the different chemical entities are different from each other. The method according to one paragraph.

Further features and aspects of the present invention will be disclosed in more detail in the following description, especially in combination with the dependent claims. Therein, each feature may be achieved in a single manner, as well as those skilled in the art, as well as in any feasible combination. Embodiments are schematically shown in the drawings. In which, the same reference numbers in these figures refer to the same or functionally identical elements.
FIG. 1 shows a schematic diagram of an analytical system and method for tracking sample identity during a process in an analytical system. FIG. 2 schematically shows the constitution of chemical substances. FIG. 3 shows the results of four example chromatographic separations from the general types n ^* (polar) and m ^* (lipophilic) analyzed by HPLC-PDA. FIG. 4 shows four example online mass spectrometric scans from the general types n ^* (polar) and m ^* (lipophilic) analyzed by Q-Tof mass spectrometry.

  FIG. 1 shows a schematic diagram of an analytical system 100 and a method for tracking sample identity during a process in the analytical system 100. Referring to FIG. 1, an example of an analytical system 100 is described. The analysis system 100 may be a clinical diagnostic system. The analysis system 100 includes at least a liquid chromatograph 102 and a mass spectrometer 104. In the current aspect, the analysis system 100 further includes a sample preparation station 106 for automated pretreatment and preparation of a sample 108, each sample containing at least one analyte of interest.

  Liquid chromatograph 102 includes a liquid chromatography separation station 110 that includes a plurality of liquid chromatography channels 112. Liquid chromatography channels 112 can also be different from one another, where there are shorter cycle time liquid chromatography channels 112 and longer cycle time liquid chromatography channels 112. However, the liquid chromatography separation station 110 may include only a plurality of faster liquid chromatography channels 112 or only a plurality of slower liquid chromatography channels 112. The analysis system 100 further includes a sample preparation / liquid chromatography interface 114 for inputting the prepared sample to any one of the liquid chromatography channels 112.

  Analysis system 100 further includes a liquid chromatography / mass spectrometer interface 116 for coupling liquid chromatography separation station 110 to mass spectrometer 104. The liquid chromatography / mass spectrometer interface 116 includes an ionization source 118 and an ion mobility module 120 between the ionization source 118 and the mass spectrometer 104. The ion mobility module 120 may be a high-field asymmetric waveform mobility mobility spectroscopy (FAIMS) module. The mass spectrometer 104 is a tandem mass spectrometer, and in particular a triple quadrupole mass spectrometer, capable of multiple reaction monitoring (MRM).

  The assay system 100 includes a predefined sample preparation workflow, each including a predefined sequence of sample preparation steps and requiring a predefined time for completion, depending on the analyte of interest. And further includes a controller 122 programmed to assign the sample 108. The controller 122 allocates (preserves) the liquid chromatography channel 112 for each preparation, depending on the analyte of interest, and based on the expected elution time, the non-overlapping LC eluate output sequences E1-. At n, it is further programmed to schedule a liquid chromatography channel input sequence I1-n for inputting the prepared sample, which allows the analyte of interest to elute from the different liquid chromatography channels 112. The controller 122 is further programmed to set and start the sample preparation start sequences S1-n, which produce the prepared sample output sequences P1-n, which match the liquid chromatography channel input sequences I1-n.

  In FIG. 1, each sample of the sample preparation start sequence S1 to n, each prepared sample output sequence P1 to n, each prepared sample of the liquid chromatography channel input sequence I1 to n, and each liquid of the liquid chromatography eluate output sequence E1 to n. The chromatographic eluate is shown in segments of the sequence that include non-overlapping adjacent segments, each segment schematically showing one reference period. Each sequence is therefore a reference period or sequence of time units, the length of which may be fixed and which remains constant over different sequences.

  The sample preparation in the sample preparation start sequence S1-n is separated by the frequency of one sample per reference period or by one or more reference periods indicated by empty segments in the sequence where sample preparation is not started. Will be started at specified intervals.

  Also, the sample preparation in the prepared sample output sequences P1-n is one or more references, indicated by the frequency of one prepared sample per reference period or by an empty segment in the sequence where the sample preparation is not complete. Complete at intervals separated by time periods.

  Preparative samples were also separated by the frequency of one liquid chromatography channel input per reference period or by one or more reference periods indicated by empty segments in the sequence with no liquid chromatography channel input. At intervals, each assigned liquid chromatography channel 112 is input according to the liquid chromatography channel input sequence I1-n.

  Also, the liquid chromatography eluate output in liquid chromatography eluate sequences E1-n is indicated at the frequency of one liquid chromatography eluate per reference period or by an empty segment in the sequence where no liquid chromatography eluate is output. It is output at intervals separated by one or more reference periods.

  Alternatively, liquid chromatography channel 112 can be coupled to liquid chromatography / mass spectrometer interface 116, and controller 122 inputs liquid chromatography eluent output to ionization source 118 one liquid chromatography eluate at a time. The valve switching 124 is controlled according to the sequence E1 to n. In particular, the liquid chromatographic eluate in the liquid chromatographic eluate output sequences E1-n may be one or more at the frequency of one liquid chromatographic eluate per reference period or according to the liquid chromatographic eluate output sequences E1-n. Inputs are made to the ionization source 118 at intervals separated by more reference periods.

  The ionization source 118 is a dual ionization source that includes an ESI source 126 and an APCI source 128, where the ionization source 118 is dependent on and contained in the liquid chromatography eluate in the liquid chromatography eluent output sequences E1-n. Depending on the analyte (s) of interest being made, the controller 122 can also select the one of the two ionization sources 126, 128 that is most appropriate. When setting the sample preparation start sequence S1-n, the controller 122 groups the samples together (also according to the ionization sources 126, 128) so that frequent switching between the ionization sources 126, 128 is prevented. They may be arranged adjacent to each other in the sequence). Ionization source switching may be planned, for example, during one or more empty reference periods.

  Continuing to refer to FIG. 1, a method for tracking sample identity during a process in analytical system 100 is described. The method includes the step of providing a sample 108. A composition 130 of different chemicals 132 is added to the sample 108 at a concentration above the detection level of the mass spectrometer 104. The composition 130 of different chemicals 132 may be added to the sample 108 before, during, or after preparation of the sample 108. In any case, the composition 130 of different chemicals 132 is added to the sample 108 before inputting the sample 108 to the liquid chromatograph 102. In this aspect, a composition 130 of different chemicals 132 is added to the sample 108 during the preparation of the sample 108. That is, a composition 130 of different chemicals 132 is added to the sample 108 at the sample preparation station 106.

  The sample 108 is then processed by the analysis system 100 along with the composition 130. In other words, the sample 108 thus prepared, together with the composition, is input to the liquid chromatograph 102, passed through the liquid chromatograph 102, and input to the mass spectrometer 104. A mass spectrometer detects and measures single and / or multiple components of sample 108. Further, composition 130 or portions of composition 130 of different chemicals 132 are detected and measured by mass spectrometer 104. The sample 108 is identified based on the detection of the substance detection signal pattern of the mass spectrometer 104, which contains substance detection signals corresponding to the composition 130 of the different chemicals 132. In other words, the signal of the mass spectrometer 104 includes not only the detection result corresponding to the sample 108, but also the signals unique to the different chemicals 132, which can unambiguously identify the sample 108. It will be possible.

  Specific details of the chemical substance 132 are described herein below. Basically, the sample contains at least one analyte of interest, and the chemical 132 is compared to the analyte of interest because the chemicals used must follow the same workflow as the analyte. It is a chemical isotope and / or has a polarity similar to the analyte of interest to ensure that it exhibits nearly identical behavior during sample processing and chromatography. The chemical substances 132 have the same molecular weight. The chemical substance 132 is at least one element selected from the group consisting of: peptides, derivatives of peptides, peptide nucleic acids, oligonucleotides, and oligomers. Further, the chemicals are designed to prevent retention on the stationary phase of the liquid chromatograph 102. This is accomplished by adapting the polarity and molecular weight of the chemical 132 to the stationary phase of the liquid chromatograph 102 to prevent the chemical 132 from being retained on the stationary phase of the liquid chromatograph 102. It is possible. For example, the polarity of the chemical 132 is adapted by using a mixture of isobaric oligomers or isotopes of the chemical 132, and the molecular weight is adapted by using a homologue, derivative or isotope label of the chemical 132. Let More specifically, the polarity of the chemistry 132 is adapted by using a mixture of oligomers or derivatives of the chemistry 132. Each oligomeric component has a labeling group 134 for molecular weight coding, which is measured intact or during the mass spectrometry process, as described in more detail herein below. Emitted and measured by mass spectrometer 104.

FIG. 2 schematically shows one configuration of the chemical substance 132. The chemical substance 132 includes a polar molecular group 136 and a lipophilic molecular group 138. The chemical substance 132 is (polar) _n- (lipophilic) _m , where (polar) indicates the polar molecular group 136, (lipophilic) indicates the lipophilic molecular group 138, and n and m are 0 or less. It is formed of a monomer block having a structure of a large integer. More specifically, the chemicals are monomer building blocks that polymerize randomly, n ^* (polar) and m ^* (lipophilic), where (polar) indicates a polar molecular group 136 (lipophilic). Represents a lipophilic molecular group 138, and n and m are integers greater than 0. In the example shown in FIG. 2, the chemical substance 132 comprises four polar molecular groups 136 followed by four lipophilic molecular groups 138. Thus, each chemical 132 includes a isobaric oligomer group 140 and at least one molecular weight coding group 142, where the isobaric oligomer group 140 and the molecular weight coding group 142 of different chemicals 132 are different from each other.

  The chemical 132 may thus be used for chemical coding of the sample. The number of target analytes is highly dependent on the application menu on analytical system 100. The requirement for the coding chemistry is therefore to include the full polar range of the application menu. This is accomplished by the composition of complex functional entities of the coding chemistry, as described above. The monomers are designed to have the same molecular weight (isobaric building blocks), so the overall molecular weight of this oligomer subunit is independent of the ratio n / m that produces the isobaric oligomer backbone. It is constant. The n and m sequence distributions are random, depending on the desired readout of the mass spectrometer 104, the molecular weight solely by using the selected ion monitor or by the molecular weight readout, and the sequence by the fragmentation pattern (daughter ion scan). It may or may not be random. Attached to this building block is a molecular weight distinguishing label. The function of the molecular weight coding group 142 is to allow the production of a wide variety of coding materials with different molecular weights. This is generated, for example, by using different amounts of stable isotope labels or different chemical functionalities within the molecular weight coding group 142. The function of the oligomer building blocks is to produce polar leaders with different chromatographic properties (logP) of the subunits but the same molecular weight. Thus, the combination of the molecular weight coding group 142 and the isobaric oligomer backbone can generate the chemical coding entity. Examples of monomers used to form the isobaric oligomer backbone include serine-propyl ether having a molecular weight of 129.16, homoserine-ethyl ether having a molecular weight of 129.16, and a molecular weight of 129.11. It is glutamic acid.

  Thus, finally, each molecular weight coding group 142 is linked to a mixture of isobaric oligomeric backbones containing the entire chromatographic space of application. Thus, independent of the analyte polarity of interest (logP), at least one member of each chemical coding entity or substance 132 is detected by the mass spectrometer 104 in the same time window as the analyte of interest. Will For example, by using seven different molecular weight coding groups 142 attached to the same isobaric oligomer backbone, it is possible to generate matrices of 107 different chemical codes. In this case, the only distinguishing criterion is molecular weight (SIM reading only).

  As mentioned above, the liquid chromatography separation station 110 includes a plurality of liquid chromatography channels 112. Therefore, it is possible to process a plurality of samples 108 at the same time. Thus, the method provides a plurality of samples 108 and adds a composition 130 of different chemicals 132 to each of the plurality of samples 108, wherein the compositions 130 added to the plurality of samples 108 are different from each other. Based on the detection of the substance detection signal pattern of the mass spectrometer 104, which has been processed by the analysis system 100 with the composition 130 of the chemical substance 132 and which contains the substance detection signals corresponding to the different compositions 130 of the chemical substance 132. , A step of identifying each of the plurality of samples 108 may be included. In this case, the number of compositions 130 may correspond to the number of liquid chromatography channels 112.

  Detailed examples of the disclosed methods are provided hereinafter.

Four different examples 1-4 of the general types of chemicals n ^* (polar) and m ^* (lipophilic) were synthesized. The following chemicals 132 with the same molecular mass (± 1 Da) were selected. Chemical substances sorted in descending order of polarity (lOgP), with hydrophobicity first: ^** Hse ^** Hse ^** Hse ^* L ^* Hse ^** Hse ^** Hse ^**- OH ^{* ; * Hse ** Hse * LE *} Hse ** Hse ** -OH *; ** Hse ** Hse * ELE * Hse ** Hse ** -OH *; ** Hse * E * Hse * ELE * Hse * ^* -OH ^* . The respective abbreviations are Hse = homoserine ethyl ether, L = leucine, LE = leucine-glutamic acid, ELE = glutamic acid-leucine-glutamic acid, E = glutamic acid.

  As an expected result of this experiment, the molecular weight should be the same for all different chemicals (± 1 Da), while the difference in polarity should result in different retention times on the chromatographic system.

Experimental method:
Materials were individually prepared by weighing 0.5 mg of each material and diluting with 1 ml of water containing 10% (v / v) acetic acid. The sample thus prepared was injected into an HPLC system (Waters Authority) equipped with a Vydac 218TP C18 column (5 μm, 2.1 × 150 mm). The following solvents were used to maintain the pump in gradient mode.

Solvent A: Water + 2% acetonitrile + 0.1% formic acid (v / v)
Solvent B: acetonitrile + 0.1% formic acid (v / v)
The following gradient was applied:
0 minutes (98% A), 15 minutes (35% A), 17 minutes (0% A), 25 minutes (0% A), 25.1 minutes (98% A), 30 minutes (98% A).

  Waters Quality PDA detector (190 nm-450 nm) and Waters Q- in positive ion electrospray ionization ion mode (scan 90-1500 Da, 30V cone voltage, 5V collision energy, 120 ° C source temperature, and 300 ° C desolvation temperature). Samples were analyzed by on-line coupling of HPLC using a Tof Premier Mass spectrometer. The scan rate was 2 Hz for the mass spectrometer and 10 Hz for the PDA detector.

Figure 3 shows the results of four example chromatographic separations from common types n ^* (polar) and m ^* (lipophilic) analyzed by HPLC-PDA. The x-axis 144 shows the retention time respectively and the y-axis 146 shows the respective retention signal respectively. Graphs 148, 150, 152, 154 show the results of the chromatographic separations of four Examples 1-4 in this order, where the results of Example 1 are provided at the bottom and the results of Example 4 are shown at the top. , Examples 2 and 3 are shown between the bottom and the top.

As ^{expected, ** Hse ** Hse ** Hse *} L * Hse ** Hse ** Hse ** -OH *; * Hse ** Hse * LE * Hse ** Hse ** -OH *; * Four different samples consisting of ^* Hse ^** Hse ^* ELE ^* Hse ^** Hse ^**- OH ^* ; ^** Hse ^* E ^* Hse ^* ELE ^* Hse ^**- OH ^* have different retention times and the reverse of the above. Elution on a phase chromatographic system, and thus for Examples 1-4, in this order, at different time points, with different polarities, i.e. different logP, as can be obtained from the respective signal peaks 156, 158, 160, 162. showed. In particular, Example 1 illustrated by graph 148 has the minimum retention time, followed by Example 2 illustrated by graph 150, followed by Example 3 illustrated by graph 152. Example 4, shown in graph 154, has the maximum retention time.

FIG. 4 shows on-line mass spectrometer scans of four Examples 1-4 from common types n ^* (polar) and m ^* (lipophilic) analyzed by Q-Tof mass spectrometry. The x-axis 164 shows the molecular mass respectively and the y-axis 166 shows the molecular mass percentage of the sample respectively. Graphs 168, 170, 172, 174 show the results of the molecular mass distribution of four Examples 1-4 in this order, where the results of Example 1 are provided at the bottom and the results of Example 4 at the top. Shown, Examples 2 and 3 are shown between the bottom and the top.

As ^{expected, ** Hse ** Hse ** Hse *} L * Hse ** Hse ** Hse ** -OH *; * Hse ** Hse * LE * Hse ** Hse ** -OH *; * ^{* Hse ** Hse * ELE * Hse} ** Hse ** -OH *; ** Hse * E * Hse * ELE * Hse ** -OH * 4 different sample of the in in this order, examples 1-4 , Have the same height and represent the same molecular weight of the pseudomolecular ion [M + H] + at 906 Da (± 1 Da), as can be obtained from the respective signal peaks 176, 178, 180, 182.

100 Analytical System 102 Liquid Chromatograph 104 Mass Spectrometer 106 Sample Preparation Station 108 Sample 110 Liquid Chromatography Separation Station 112 Liquid Chromatography Channel 114 Sample Preparation / Liquid Chromatography Interface 116 Liquid Chromatography / Mass Spectrometer Interface 118 Ionization Source 120 Ion Mobility Module 122 Controller 124 Valve switching 126 ESI source 128 APCI source 130 Composition 132 Chemical substance 134 Labeling group 136 Polar molecular group 138 Lipophilic molecular group 140 Isobaric oligomer group 142 Molecular weight coding group 144 Hours 146 Retention signal 148 For chromatographic separation of Example 1 Results 150 Chromatographic separation results for Example 2 152 Chromatographic separation results for Example 3 154 Chromatographic separation results for Example 4 156 Signal peak for Example 1 158 Signal peak for Example 2 160 Signal peak for Example 3 162 Signal peak for Example 4 164 Molecular mass 166 Molecular mass percentage 168 Molecular mass distribution for Example 1 170 Molecular mass distribution for Example 2 172 Molecular mass distribution for Example 3 174 Molecular mass distribution for Example 4 176 Signal peak for Example 1 178 Signal peak for Example 2 180 For Example 3 Signal peak 182 Signal peak for Example 4

Claims (15)

A method for tracking sample identity during a process in an analytical system (112), the analytical system (112) including a liquid chromatograph (102) and a mass spectrometer (104), the method comprising:
-Providing a sample (108),
Adding a composition (130) of different chemicals (132) to the sample (108) at a concentration above the detection level of the mass spectrometer (104),
Processing the sample (108) with the composition (130) by the analytical system (112),
-Detecting and / or measuring one or more components of the sample (108) by a mass spectrometer (104),
-A composition (130) or a portion of a composition (130) of different chemicals (132) is detected and measured by a mass spectrometer (104),
-Identifying the sample (108) based on the detection of the substance detection signal pattern of the mass spectrometer (104), which comprises a substance detection signal corresponding to the composition (130) of the chemical substance (132), Method.   The method of claim 1, wherein the composition (130) of different chemicals (132) is added to the sample (108) before, during or after the preparation of the sample (108).   The method of claim 1 or 2, wherein the chemical (132) is designed to prevent the chemical (132) from being retained on the stationary phase of the liquid chromatograph (102).   The polarity and molecular weight of the chemical (132) are adapted to the stationary phase of the liquid chromatograph (102) so as to prevent the chemical (132) from being retained on the stationary phase of the liquid chromatograph (102). The method of claim 3, wherein the method is:   The method of claim 4, wherein the polarity of the chemical (132) is adapted by using a mixture of isobaric oligomers or isotopes of the chemical (132).   Method according to claim 3 or 4, wherein the molecular weight is adapted by using homologues, derivatives or isotopic labels of the chemical substance (132).   The method of claim 4, wherein the polarity of the chemical (132) is adapted by using a mixture of oligomers or derivatives of the chemical (132).   Each oligomeric component has a labeling group (134) for molecular weight coding, which is measured intact or released during the mass spectrometric process, and the mass spectrometer (104). 8.) The method according to claim 7, which is measured by   9. The sample of claim 1-8, wherein the sample (108) comprises at least one analyte of interest, the chemical (132) is a chemical isotope, and / or has a polarity similar to the analyte of interest. The method according to any one of claims.   10. The method of any one of claims 1-9, wherein the chemical (132) comprises polar molecular groups (136) and lipophilic molecular groups (138). The chemical substance (132) is (polar) _n- (lipophilic) _m , wherein (polar) indicates a polar molecular group (136), (lipophilic) indicates a lipophilic molecular group (140), and 11. The method according to claim 10, which is formed from a monomer block having a structure in which n and m are integers greater than 0. The chemical substance (132) is a monomer building block that randomly polymerizes, n ^* (polar) and m ^* (lipophilic), in which (polar) represents a polar molecular group (136), and (lipophilic) Represents a lipophilic molecular group (138), and n, m are integers greater than 0.   A method according to any one of claims 1 to 12, wherein the liquid chromatograph (102) comprises a liquid chromatography separation station (110) comprising a plurality of liquid chromatography channels (112), the method further comprising: A composition (130) that provides a plurality of samples (108) and adds a composition (130) of different chemicals (132) to each of the plurality of samples (108), where the compositions (130) are added to the plurality of samples (108). A plurality of different samples (108), together with the composition (130) of the chemical substance (132), processed by the analytical system (112) and corresponding to the different composition (130) of the chemical substance (132). The method, comprising identifying each of the plurality of samples (108) based on detection of the substance detection signal pattern of the mass spectrometer (104) including the detection signal.   14. The method of claim 13, wherein the number of compositions (130) corresponds to the number of liquid chromatography channels (112).   Each chemistry (132) comprises an isobaric oligomeric group (140) and at least one molecular weight coding group (142), wherein the isobaric oligomeric group (140) and molecular weight coding group of a different chemistry (132). 15. A method according to any one of claims 1-14, wherein (142) are different from each other.

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